Helminths in horses: use of selective treatment for the control of strongyles

The current level of anthelmintic resistance in the horse-breeding industry is extremely high and therefore more emphasis is being placed on studies that focus on the judicious use of anthelmintic products. The aims of the study were to: 1) establish if there is variation in the egg excretion pattern of strongyles between the different age classes of Thoroughbred horses in the Western Cape Province (WCP), 2) test if a selective treatment approach successfully reduces the number of anthelmintic treatments and maintains acceptably low helminth burdens in adult Thoroughbred horses, and 3) evaluate the efficacy of subsampling large horse herds for faecal egg counts (FECs) to monitor the strongyle burden. In 2001 the FECs of 4 adult mare, 5 yearling and 3 weanling herds from 8 different farms were compared in the WCP Within the mare herds there were generally fewer egg-excreting individuals with lower mean FECs compared with the younger age classes. Individual faecal samples were collected every 3-4 weeks from 52 adult Thoroughbred mares from 1 farm in the WCP during a 12-month period (2002/2003). Animals with strongyle FECs > or =100 eggs per gram (epg ) were treated with an ivermectin-praziquantel combination drug (Equimax oral paste, Virbac). The mean monthly strongyle FEC for the entire group was <300 epg throughout the study and the number of treatments was reduced by 50 %. Resampling methods showed that an asymptote to mean FEC was reached at 55 animals for each of the pooled weanling, yearling and mare egg counts. Resampling within 4 different mare herds recorded asymptotes of between 24 and 28 animals. Subsampling entire herds for FECs therefore provided an effective approach to treatment management. This study demonstrates that selective treatment is both a practical and an effective approach to the management of anthelmintic resistance.     

A survey of benzimidazole resistant nematodes in sheep in three countries of southern England.

Faecal samples were collected from 209 randomly selected sheep farms in southern England, and the nematode eggs extracted were tested by an egg hatch assay and by a larval development test for their resistance to benzimidazoles. Benzimidazole resistance was found on 35 per cent of farms tested in East Sussex, 44 per cent in Oxfordshire and 61 per cent in West Sussex.     

An introductory survey of helminth control practices in south africa and anthelmintic resistance on Thoroughbred stud farms in the Western Cape Province

Fifty-one per cent of 110 questionnaires, designed for obtaining information on helminth control practices and management on Thoroughbred stud farms in South Africa, were completed by farmers during 2000. The number of horses per farm included in the questionnaire survey ranged from 15 to 410. Foals, yearlings and adult horses were treated with anthelmintics at a mean of 7.3 +/- 3.0, 6.6 +/- 2.7 and 5.3 +/- 2.3 times per year, respectively. An average of 3.4 different drugs were used annually, with ivermectin being used bymost farmers during 1997-2000. On 43% of farms the weights of horses were estimated by weigh band and 45% of farmers estimated visually, while both were used on 7% of farms and scales on the remaining 5%. Doses were based on average group weight on 50% ofthe farms and on individual weights on 46%. Forty-three per cent of farmers performed faecal egg count reduction tests (FECRT). Most farmers rotated horses between pastures and treated new horses at introduction. Faecal removal was practiced on 61% of farms and less than 50% of farmers used alternate grazing with ruminants. Faecal egg count reduction tests were done on 283 horses, using oxibendazole, ivermectin and moxidectin on 10,9 and 5 farms, respectively, in the Western Cape Province during 2001. While the efficacy of oxibendazole was estimated by FECRT to range from 0-88% and moxidectin from 99-100%, ivermectin resulted in a 100% reduction in egg counts. Only cyathostome larvae were recovered from post-treatment faecal cultures.     

Benzimidazole resistance in equine cyathostomes in Slovakia

The present study included 19 stud farms, including 243 horses, that were investigated for the occurrence of anthelmintic resistant cyathostomes. The number of horses on the farms varied from nine to more than 100, and horses of all ages were included. A minimum of seven horses were used for faecal egg count reduction (FECR) tests. The anthelmintics included were: fenbendazole (paste formulation), ivermectin (paste formulation) and pyrantel (powder). Resistance to benzimidazoles was detected on 14 farms, with FECR values ranging from 65.1 to 86.3%. Larval cultures after fenbendazole treatment revealed exclusively cyathostome larvae. Ivermectin was tested on eight farms and proved to be effective on all. Pyrantel was tested on two farms and FECR test indicated high efficacy (92-97%). Egg hatch assay (EHA) results showed that mean concentrations of thiabendazole that inhibited hatching in 50% of the eggs (ED(50)) in resistant populations were over 0.1 microg ml(-1). The results of our study suggest widespread resistance to fenbendazole in equine cyathostomes in Slovakia, and possible strategies to delay anthelmintic resistance are discussed briefly.     

A Field Study on the Effect of Some Anthelmintics on Cyathostomins of Horses in Sweden

The objective of the study was to investigate different aspects on the efficacy of three anthelmintics on cyathostomin nematodes of Swedish horses. A faecal egg count reduction (FECR) test was performed on 26 farms. Horses were treated orally with recommended doses of ivermectin, pyrantel pamoate or fenbendazole. Faecal samples were collected on the day of deworming and 7, 14 and 21 days later. No resistance was shown against ivermectin; the FECR was constantly >99%. The effect of pyrantel was assessed as equivocal in 6 farms 14 days after treatment; the mean FECR was 99%. As many as 72% of the fenbendazole-treated groups met the criteria for resistance; the mean FECR was 86%, ranging from 56% to 100%. A re-investigation of two farms where pyrantel resistance had been suspected clearly revealed unsatisfactory efficacy of pyrantel on one of these farms; the FECR varied from 72% to 89%. Twenty-six of the horses previously dosed with pyrantel or fenbendazole, and which still excreted ≥150 eggs per gram of faeces 14 days after treatment, were dewormed with ivermectin and fenbendazole or pyrantel in order to eliminate the remaining cyathostomins. A total of 13 cyathostomin species were identified from horses that initially received fenbendazole and seven species were identified from pyrantel-treated individuals. The egg reappearance period (ERP) following treatment with ivermectin and pyrantel was investigated on two farms. The shortest ERP after ivermectin treatment was 8 weeks and after pyrantel was 5 weeks. We conclude that no substantial reversion to benzimidazole susceptibility had taken place, although these drugs have scarcely been used (<5%) in horses for the last 10 years. Pyrantel-resistant populations of cyathostomins are present on Swedish horse farms, but the overall efficacy of pyrantel is still acceptable.     

Anthelmintic resistance and sheep management practices in south western Victoria

Twenty-eight farms in 7 shires in south western Victoria were selected and tested for presence of benzimidazole-resistant nematodes between November 1979 and June 1981. Mean faecal egg counts of sheep were less than 100 strongyloid eggs/g on 11 farms. Faecal egg count reduction tests were conducted on the remaining 17 farms and thiabendazole was less than 90% efficient in reducing egg counts in sheep from 5 (29%) of these farms. Thiabendazole-resistant Teladorsagia circumcincta were identified at necropsy of experimentally infected treated sheep. In further studies a survey of 104 farms was conducted in the Mount Rouse and Dundas shires of western Victoria in 1981 and 1982 respectively to determine the prevalence of anthelmintic resistance in these shires. Mean faecal egg counts among weaner sheep in the winter-spring of both years were less than 100 eggs/g which indicated low levels of parasitic nematode populations. A faecal egg count reduction test was conducted on 10 farms and thiabendazole was less than 90% efficient on 3; levamisole was greater than 90% efficient in all 10 tests. Most of the surveyed farms carried Merino or Merino crossbred sheep at 10 to 15 dry sheep equivalents per ha and weaners were treated with anthelmintics 3 to 6 times per year. Management procedures based mainly on anthelmintic therapy were effective in controlling nematode populations in weaner sheep, although many producers alternated between different groups of anthelmintics within the same year contrary to current recommendations for long-term preservation of anthelmintic efficacy. It was concluded that anthelmintic resistance was not of practical importance to the majority of sheep producers in the region.     

Prevalence of gastrointestinal nematodes in dairy heifers in western Canada

A survey was conducted to estimate the prevalence of gastrointestinal nematodes in grazing dairy replacement heifers in western Canada. Fecal samples were collected from 566 animals on 34 farms distributed over the three prairie provinces. Fecal egg counts were performed using a modified Wisconsin centrifugation technique. The overall mean farm fecal egg count was 16.0 eggs per gram feces (range 0.7-78.1 epg). Gastrointestinal nematodes were demonstrated on every farm examined, and 93.3% of all individuals tested were positive.     

Nematode control practices and anthelmintic resistance in dairy calves in the south west of England.

A postal survey of worming practices on West Country dairy farms was undertaken and farmers were requested to send faecal samples for nematode egg counts. The majority of the farmers who responded had a nematode control policy which was based on a mixture of anthelmintics and pasture rotation. Sixty-five per cent turned out calves on to the same paddock each year and 57 per cent treated their stock with anthelmintics during the second year. Ninety farms submitted samples for analysis but only 16 samples contained sufficient eggs to justify repeat egg counts and only eight of these produced enough eggs for in vitro tests. The small number of positive samples, even into the latter part of the year suggests a heavy use of anthelmintics with relatively clean pasture. One Somerset farm had larvae which developed in high concentrations of ivermectin, and eggs were still being passed after two treatments with ivermectin at the manufacturer's recommended dose seven days apart. Of 100 male nematodes isolated from two of the calves, 88 were Cooperia species and 12 were Nematodirus species. A controlled trial with this isolate in eight Friesian male calves showed a 44 per cent reduction in egg counts at day 7 but no significant reduction in worm burden postmortem. This is the first reported case of ivermectin-resistant Cooperia species in cattle in the United Kingdom.     

Comparative use of faecal egg count reduction test,egg hatch assay and beta-tubulin codon 200 genotyping in small strongyles (cyathostominae) before and after benzimidazole treatment

A survey on benzimidazole (BZ) resistance in small strongyles was performed on three farms in the tenth region in Chile. Samples from a total of 100 horses were tested using the faecal egg count reduction test (FECRT), the egg hatch assay (EHA) and an allele-specific PCR for the detection of beta-tubulin isotype 1 genes coding for phenylalanine (phe) or tyrosine (tyr) at codon 200. In the past, BZ-type drugs have been used within anthelmintic campaigns on all the three farms. This has predictably led to a high degree of BZ resistance at the Valdivia and Ri?ihue farms and to a lesser degree at the Frutillar farm, as demonstrated by all the three tests. The FECRT indicated resistance in every farm by faecal egg count reductions (FECR) of 27% (S.D. +/- 33), 26.5% (S.D. +/- 26.9) and 83.9% (S.D. +/- 22.8) for the Valdivia, Ri?ihue and Frutillar farms, respectively. With the EHA, the following mean LD(50) values were found before and after treatment with fenbendazole (FBZ): 0.093, 0.141 and 0.066 microg TBZ/ml and 0.149, 0.158 and 0.091 microg TBZ/ml, respectively, for the Valdivia, Ri?ihue and Frutillar samples. The corresponding LD(96) values were 0.222, 0.263 and 0.188 microg TBZ/ml before treatment and 0.316, 0.322 and 0.221 microg TBZ/ml after treatment, indicating BZ resistance in all the cases. Genotyping was performed on more than 1700 single larvae, at least 10 per faecal sample, for 98 pre- and 66 post-treatment samples. Despite a general trend toward higher percentages of phe/tyr and tyr/tyr individuals following treatment, no statistically significant difference was found between these two and the phe/phe genotype percentages. However, a significantly negative correlation was detected between the LD(50) values and the phe/phe percentages and there was a positive correlation between the FECRT results and the phe/phe percentages. Thus, there seems to be a difference in the significance of the codon 200 polymorphism in the mechanisms of BZ resistance in small strongyles of the horse and sheep trichostrongyles.     

The use of age-clustered pooled faecal samples for monitoring worm control in horses

A study was performed on two horse farms to evaluate the use of age-clustered pooled faecal samples for monitoring worm control in horses. In total 109 horses, 57 on farm A and 52 on farm B, were monitored at weekly intervals between 6 and 14 weeks after ivermectin treatment. This was performed through pooled faecal samples of pools of up to 10 horses of the groups 'yearlings' (both farms), '2-year-old' (two pools in farm A), '3-year-old' (farm A) and adult horses (four pools on farm A and five pools on farm B), which were compared with the mean individual faecal egg counts of the same pools. A very high correlation between the faecal egg counts in pooled samples and the mean faecal egg counts was seen and also between the faecal egg counts in pooled samples and larval counts from pooled faecal larval cultures. Faecal egg counts increased more rapidly in yearlings and 2-year-old horses than in older horses. This implied that in these groups of young animals faecal egg counts of more than 200 EPG were reached at or just after the egg reappearance period (ERP) of 8 weeks that is usually indicated for ivermectin. This probably means that, certainly under intensive conditions, repeated treatment at this ERP is warranted in these young animals, with or without monitoring through faecal examination. A different situation is seen in adult animals. Based on the mean faecal egg counts on both farms and on the results of pooled samples in farm A, using 100 EPG as threshold, no justification for treatment was seen throughout the experimental period. However, on farm B values of 100 EPG were seen at 9 and 11, 13 and 14 and 14 weeks after ivermectin treatment in pools 10, 12 and 13, respectively. This coincided with the presence of one or two horses with egg counts above 200 EPG. The conclusion is that random pooled faecal samples of 10 adult horses from a larger herd, starting at the ERP and repeating it at, for instance, 4-week intervals, could be used for decisions on worm control. However, there would be a certain risk for underestimating pasture contamination through missing high-egg excreters. An alternative use of pooled samples would be as a cheap first screening to detect which adult horses really contribute to pasture contamination with worm eggs on a farm. All horses should be sampled and subsequently animals from 'positive' pools can be reexamined individually.     

Serological surveillance of equine viral arteritis in the United Kingdom since the outbreak in 1993

Serological analysis of blood samples submitted to the Animal Health Trust showed that during 1995, 185 of 9203 unvaccinated horses (2.0 per cent) tested positive for antibodies to equine arteritis virus (EAV), and that during 1996, 46 of 8851 unvaccinated horses (0.52 per cent) tested positive. During both years thoroughbreds were the predominant breed tested and only a small proportion of these (<0.3 per cent), consisting predominantly of imported mares, were seropositive. In contrast, among standardbred horses, from which samples were actively solicited in 1995, 84 of 454 (18.5 per cent) were seropositive. Among standardbreds there was a difference in prevalence between types of horses, with 3.7 per cent of racing horses, 25 per cent of non-racing horses and 41 per cent of stallions testing seropositive. Investigations of seropositive stallions identified during 1994 and 1995 demonstrated that clinically inapparent equine viral arteritis (EVA) had occurred previously in the UK. Of 50 seropositive stallions, nearly half were standardbreds and nearly all had been imported from either North America or the European Union. Whether 34 seropositive stallions were shedding virus in their semen was established either by test mating, by the serology of the covered mares, or by investigation by MAFF following the introduction of the Equine Viral Arteritis Order 1995. Nine of the stallions (26.5 per cent) were identified as presumptive shedders of EAV in semen and among specific breeds, viral shedding was identified in six of 15 (40 per cent) standardbreds and three of nine (33 per cent) warmbloods. In contrast with the outbreak of EVA in the UK in 1993, no signs of disease typical of EAV infection were reported during these investigations, even in mares test mated to stallions shedding the virus.     

Cases of reduced cyathostomin egg-reappearance period and failure of Parascaris equorum egg count reduction following ivermectin treatment as well as survey on pyrantel efficacy on German horse farms

In 2003 and 2004, on a total of 63 different German horse farms, a survey using the faecal egg count reduction (FECR) test was performed to investigate the efficacy of ivermectin (IVM, Ivomec) and pyrantel (PYR, Banminth) treatment against gastro-intestinal nematodes in a total of 767 horses. IVM treatment resulted in 100% reduction of the cyathostomin egg production 14 and 21 days post-treatment (d.p.t.) on 37 farms. On the remaining five farms, the mean faecal egg count reduction ranged between 97.7 and 99.9%. The mean cyathostomin FECR following PYR treatment ranged between 92.2 and 100% on the 25 farms tested. Therefore, based on the 90% FECR threshold suggested for detection of anthelmintic resistance in horses, neither IVM nor PYR anthelmintic resistance was detected. However, if the thresholds recommended for the detection of resistance in small ruminants were applied, on one and four farms signs of reduced IVM and PYR efficacy, respectively, were observed. In 2005, to further investigate these findings, the cyathostomin egg-reappearance period (ERP) following IVM treatment was examined on six selected farms, two of which were found to show less than 99.8% FECR in the previous survey. On these two latter farms, the ERP was less than 5 weeks, while on the other four it was at least 8 weeks. Earlier investigations described IVM cyathostomin ERP of at least 9 weeks. The efficacy of IVM to reduce Parascaris equorum egg excretion was also studied. On one farm in 2 consecutive years, IVM treatment did not lead to a significant reduction in P. equorum faecal egg counts in one and five young horses, respectively.     

Daily Variability of Strongyle Fecal Egg Counts in Horses

Strongyle parasites are ubiquitous in grazing horses and constitute a potential threat to equine health. Feces were collected from six horses four times daily over a period of 5 days. Fecal egg counts (FECs) were performed to identify any diurnal rhythms in strongyle egg shedding and to quantify variability at the different levels: individual horses, repeated counts, repeated subsamples, different time points, and different days. No significant differences in FECs were found between the different time points (P = .11). The variables—horse, day, subsample, and egg count—accounted for a variance of 104.83, 0.10, 7.24, and 5.61, respectively. The apparent lack of additional variability between the four different time points suggests that time of the day chosen for collecting fecal samples does not constitute a source of error in field studies. The majority of variability exists between different subsamples and repeated egg counts on the same subsamples, whereas the variability of FECs between following days can be considered negligible. The findings of this study have implication for designing and performing field surveillance of strongyle FEC levels and applying the FEC reduction test for evaluating anthelmintic efficacy.     

Endoparasite control management on horse farms – lessons from worm prevalence and questionnaire data

Reasons for performing study: Increasing prevalence of anthelmintic resistance in equine nematodes calls for a reexamination of current parasite control programmes to identify factors influencing control efficacy and development of resistance. Objectives: To investigate if associations occur between prevalence of parasitic nematodes and management practices. Methods: German horse farms (n = 76) were investigated in 2003 and 2004. Information on farm and pasture management with respect to endoparasite control measures obtained using a questionnaire survey. Faecal examinations were performed in parallel. Results: Horses (n = 2000) were examined by faecal nematode egg counts, grouped into foals, yearlings and mature individuals for statistical analyses. Farms were categorised into 3 types, riding, stud farms and small holdings. Count regression models were used to analyse strongyle faecal egg count data. Following dichotomisation of faecal egg count (FEC) data, prevalence of strongyle and Parascaris equorum infections were assessed by logistic regression models as a function from various management factors. Yearlings on stud farms showed a 2‐fold higher risk of being positive for strongyle FEC, higher (i.e. ≥3 per year) anthelmintic drug treatment frequencies were associated with reduced strongyle infection rates only in mature individuals but not in foals or yearlings, foals on farms fertilising pastures with horse manure had a significantly higher risk of being P. equorum FEC positive and yearlings on stud farms were more often showing incomplete FECR following anthelmintic treatment compared to yearlings on other farm types. The mean yearly treatment frequencies per age group were: foals 4.52, yearlings 3.26 and mature horses 2.72 times, respectively. Conclusion and potential relevance: To delay the development of anthelmintic, resistance management should include additional nonchemotherapeutic parasite control strategies, FEC‐monitoring, controlled quarantine treatment of new arrivals and control of efficacy by the faecal egg count reduction test on a regular basis.     

广东省部分地区马肠道寄生虫感染情况调查

【目的】调查广东省内部分地区马匹寄生虫感染情况,探索区域内马匹寄生虫感染优势虫种、感染情况和耐药情况,为区域内马肠道寄生虫病的防治提供参考。【方法】采用饱和食盐溶液漂浮法和饱和蔗糖溶液漂浮法对广东省内7个城市24个马术俱乐部375份新鲜马粪样品进行处理,光学显微镜下观察虫卵/卵囊的形态结构特征并进行分类,并根据麦氏计数法计算每克粪便中虫卵/卵囊数(EPG),统计感染率和感染强度。对EPG>500的阳性样本进行粪便虫卵减少量计数试验(FECRT),以评估各马术俱乐部流行寄生虫耐药情况。提取5份<1岁幼驹的粪便样本DNA,利用巢式PCR检测样品是否存在隐孢子虫感染。【结果】375份样品中有120份样品呈寄生虫虫卵/卵囊阳性,总感染率为32.00%。共检出4种肠道寄生虫,分别为圆线虫、马副蛔虫、类圆线虫和马蛲虫,总感染率分别为30.40%、3.70%、0.80%和0.30%。1～4岁马匹感染率显著高于其他年龄段(P<0.05),但不同性别马匹寄生虫感染情况无显著差异(P>0.05)。受调查马匹感染强度以低排卵(EPG<200)为主,占马匹总数83.47%。FEC...     

The persistence of benzimidazole-resistant cyathostomes on horse farms in Ontario over 10 years and the effectiveness of ivermectin and moxidectin against these resistant strains

Three clinical trials with fecal egg count reduction tests and coproculture were conducted on 2 standardbred farms in Ontario. On Farm A, the treatment groups were mebendazole and ivermectin in trial 1, and fenbendazole and moxidectin in another. On Farm B, treatment groups were mebendazole and ivermectin. All horses treated with mebendazole or fenbendazole were subsequently treated with ivermectin or moxidectin. Strongyle eggs/g feces were estimated pre- and post-treatment using the Cornell-McMaster dilution and Cornell-Wisconsin centrifugal flotation techniques. After treatment, there was no change in the arithmetic mean eggs/g feces for horses given mebendazole, and a reduction of only 49.1% for those given fenbendazole. All horses receiving ivermectin or moxidectin had their egg counts reduced to 0. Only cyathostomes were found on culture. On both farms the benzimidazole resistant strains appeared to have persisted for at least 10 years. Development of and monitoring for anthelmintic resistance are briefly discussed.     

Association between variation in faecal egg count for a mixed field-challenge of nematode parasites and ovine MHC-DQA2 polymorphism

The selection of sheep that are resistant to gastrointestinal parasites and have lower faecal egg counts (FECs) has been the subject of extensive research. This has led to the speculation that the Major Histocompatibility Complex (MHC) genes could be used as markers to reduce FEC. In this study, associations between variation in ovine MHC-DQA2 and various measures of FEC recorded at two times (approximately 4 and 9 months of age) were investigated in a large group of New Zealand lambs (n=4676), derived from 185 different sire-lines, of a variety of breeds and raised on 25 separate farms. Pair-sample t-tests revealed that FEC for Nematodirus spp., Strongyle spp. and total FEC differed significantly between the two assessments. A total of twenty one DQA2 alleles or DQA2-DQA2-like haplotypes were identified, with allele/haplotype presence and frequency varying significantly between farms. For example, allele *0103 was observed on all farms, ranging in frequency from 0.2 to 60.9%, while haplotype *0101-*1601 was only present on one farm, in two lambs. A number of associations between the presence/absence of these alleles and egg counts were observed, but nearly all the allelic/haplotypic associations were age and parasite specific, suggesting that immune response is both age and challenge (parasite species mix) dependent. The exception was allele *1201 which was associated with increased total FECs at both 4 and 9 months of age; with it either being, or tending toward being, significantly associated with both increased Strongyle spp. and Nematodirus spp. counts as well. However, the observed increases in egg counts were small and ranged between 5 and 32 eggs per gram. In conclusion, we believe that the MHC plays an important role in parasite resistance, but that the MHC-nematode interaction is complex and thus the development of a single gene-marker based on the "MHC effect" is unlikely.     

Searching for ivermectin resistance in Dutch horses

A study was conducted to evaluate the occurrence of resistance against, in particular, ivermectin in cyathostomins in the Netherlands. Seventy horse farms were visited between October 2007 and November 2009. In initial screening, faecal samples were collected 2 weeks after deworming with either ivermectin, moxidectin or doramectin. Pooled faecal samples from a maximum of 10 horses were examined for worm eggs using a modified McMaster technique and for worm larvae after faecal larval cultures. In total 931 horses were involved. On 15 of 70 farms eggs and/or larvae were found. On 8 of these 15 farms a FECRT with ivermectin was performed on 43 horses. Efficacy of ivermectin against cyathostomins of 93% was found in one animal on one farm. Additionally, the strategies and efforts of the horse owners to control cyathostomins, as well as risk factors for the development of macrocyclic lactone resistance were evaluated with a questionnaire. Strikingly, many responders indicated that the control of cyathostomins in horses is achieved through very frequent deworming. Fourteen percent of these owners deworm seven times per year or more. On 34% of the 70 farms treatment was repeated within the Egg Reappearance Period of a product.     

Evaluation of a larval development assay (DrenchRite) for the detection of anthelmintic resistance in cyathostomin nematodes of horses

A larval development assay (LDA, DrenchRite) was evaluated to determine the effectiveness of this method in detecting anthelmintic resistance in cyathostomin nematodes of horses. A total of 15 horse farms from Georgia and South Carolina (USA) and Population S ponies from the University of Kentucky (USA) were included in this study. Nematode eggs were extracted from pooled fecal samples and placed into the wells of a DrenchRite plate for testing against thiabendazole (TBZ), levamisole (LEV) and 2 ivermectin (IVM) analogs (IVM-1, IVM-2). After a 7-day incubation larvae in each well were counted and data were analyzed by logistic regression. Resistance status of each farm for different drugs was determined in a separate study using a fecal egg count reduction test. LDA were performed on the 15 farms once, however, the Population S cyathostomins were assayed on 3 separate occasions to estimate the consistency of results between assays. Mean TBZ LC50 for oxibendazole resistant, suspected resistant and sensitive farms were 0.2015, 0.1625, and 0.1355 microM, respectively. For LEV, mean LC50 for PYR resistant, suspected resistant and sensitive farms were 1.590, 1.8018 and 1.4219 microM, respectively. All 15 farms had worms susceptible to IVM; mean LC50 for IVM-1 and for IVM-2 were 7.5727 and 87.9718 nM, respectively. A linear mixed model was fitted to the data to determine the relationship between LC50 and LC95 and resistance status for each farm. No meaningful relations were found. Consistency of assays varied between drugs, being best for TBZ and worst for LEV and IVM-1. All farms in this study had benzimidazole-resistant nematodes; therefore usefulness of DrenchRite for discriminating susceptibility versus resistance to this drug class could not be accurately assessed. Moreover, since all farms tested were sensitive to IVM and resistance to this drug class has not yet been reported in cyathostomins, it is not possible to assess accurately the usefulness of DrenchRite LDA for detecting IVM resistance at this time. Assay results for LEV suggest that LEV in a LDA does not yield data that is useful in estimating PYR efficacy in vivo. Based on results for PYR/LEV, the current high prevalence of benzimidazole resistance, no known cases of IVM resistance, and the sometimes extreme variation in results seen in many of the assays, DrenchRite LDA cannot be considered a useful tool for the diagnosis of resistance in cyathostomins of horses at present.